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Product Details
Source
Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016) is a rabbit monoclonal antibody recombinantly expressed from human 293 cells (HEK293), which provides higher batch consistency and long term security of supply. It shows superior performance in flow cytometry assays with higher affinity and better specificity.
Application
Flow Cytometry (Evaluation of cell surface expressed CARs of varying specificity containing a G4S linker within the scFv of the extracellular domain).
Clone
016
Species
Rabbit
Isotype
Rabbit IgG | Rabbit Kappa
Specificity
It can specifically recognize proteins or ScFv containing (G3S)n or (G4S)n linkers, where n = 2, 3, 4... That is, it can recognize linkers with a minimum length of 2 repeats for both (G3S)n and (G4S)n.
Conjugate
PE
Excitation Wavelength: 488 nm / 561 nm
Emission Wavelength: 575 nm
Recommended Dilution
1:50
Formulation
Lyophilized from 0.22 μm filtered solution in PBS, 0.03% Proclin 300, pH7.4, 0.2% BSA with trehalose as protectant.
Contact us for customized product form or formulation.
Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please protect from light and avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 24 months in lyophilized state;
- -70°C for 12 months after reconstitution.
- 2-8°C for 12 months after reconstitution.
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Performance Data
Bioactivity-FACS
Flow cytometric analysis of Anti-MSLN CAR-293 cells staining with PE-Labeled Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016) (Cat. No. G4S-PFMY25) at 1:50 dilution (2 μL of the antibody stock solution corresponds to labeling of 1e6 cells in a final volume of 100 µL), compared withisotype control antibody. PE signal was used to evaluate the binding activity (QC tested).
Protocol
Non-specificity of PE-Labeled Monoclonal Anti-G4S linker Antibody, Rabbit IgG (016) (Cat. No. G4S-PFMY25) binding to CD3+ cellspresent in human PBMC. 5e5 of human PBMCs were simultaneously stained with FITC-labeled anti-CD3 antibody and PE-Labeled Anti-G4S Linker Antibody (016) (2 μL of the antibody stock solution corresponds to labeling of 5e5 cells in a final volume of 100 µL) andwashed and then analyzed with FACS. Both FITC and PE positive signals was used to evaluate the non-specific binding activity to humanCD3+ cells (QC tested).
Protocol
Typical Data
Immunohistochemical analysis of paraffin embedded anti-CD19 CAR-T cells labelled with G4S-PFMY25 at 1/400 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with lHC staining protocol.
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